抑制性消减杂交分离苹果缺铁诱导的相关基因  

Isolation of Fe Deficiency-induced Genes by Suppression Subtractive Hybridization in Malus xiaojinensis

作　　者：张玉刚[1] 许雪峰 李天忠[1] 王忆[1] 孔瑾[1] 韩振海[1] 

机构地区：[1]中国农业大学园艺植物研究所

出　　处：《园艺学报》2007年第3期555-560,共6页Acta Horticulturae Sinica

基　　金：北京市重点实验室资助项目;国家自然科学基金项目(30170552)

摘　　要：以铁高效基因型植物——苹果属小金海棠(Malus xiaojinensis Cheng et Jiang)为材料,利用抑制性消减杂交(SSH)方法,成功构建了小金海棠缺铁条件下的消减cDNA文库。该文库包含1 152个克隆,通过PCR检测插入片段大小在300 ～1 000 bp之间。对构建的消减cDNA文库进行差异筛选,得到差异表达片段(ESTs) 105个。GenBank上BLAST结果如下: 50个ESTs与其它物种同源性较高,视为已知基因,并根据其功能分为9个组; 32个ESTs在GenBank中无法查到对应的同源序列,可能代表了新基因;另外23个是与已知ESTs重复的基因片段。In order to isolate genes involved in Fe nutrition, suppression subtractive hybridization （SSH） was performed using mixed cDNAs prepared from Malus xiaojinensis roots treated with Fe deficiency as tester and treated with Fe sufficiency as drivers. A forward subtractive cDNA library was constructed, from which 1 152 recombinant colonies were picked and amplified. Through differential screening of the subtractive cDNA library, 105 express sequence tags （ESTs） were identified as Fe deficiency stress induced. Of the 105 ESTs, 50 contain regions with 60% - 90% identity to their homolog in GenBank, 32 are expected to be novel genes, 23 are repeated fragments to other genes. The ESTs with significant protein homology were sorted into 9 functional categories.

关 键 词：苹果 小金海棠 抑制性消减杂交 CDNA文库 差异筛选 

分 类 号：S661[农业科学—果树学]